Review



anti psyk  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 95
      Buy from Supplier

    Structured Review

    Cell Signaling Technology Inc anti psyk
    Anti Psyk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+phospho+syk+psyk/Phospho-Syk+(Tyr525%2F526)+Rabbit+mAb/pmc12913675-361-15-16
    Average 95 stars, based on 221 article reviews
    anti psyk - by Bioz Stars, 2026-09
    95/100 stars

    Images

    Related Articles

    Expressing:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.

    Positive Control:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.

    Western Blot:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.

    Comparison:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.

    Software:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.

    Negative Control:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.

    Cell Counting:

    Article Title: ArfGAP with the SH3 Domain, Ankyrin Repeat and PH Domain 1 Inversely Regulates Programmed Death-Ligand 1 Through Negative Feedback of Phosphorylated Epithelial Growth Factor Receptor and Activation of Nuclear Factor-Kappa B in Non-Small Cell Lung Cancer
    Article Snippet: Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).Protein concentrations were determined using Bio‑Rad Protein Assay Dye Reagent Concentrate (#5000006JA; Bio‑Rad Laboratories, Inc).. The following primary antibodies were used: Anti‑phospho-epidermal growth factor receptor (pEGFR) (Cell Signaling Technology; cat. no. 2236 1:1000), anti‑PD-L1 (Cell Signaling Technology; cat. no. 13684 1:1000), anti- phospho-ACK1 (pACK1) (Cell Signaling Technology; cat. no. 3138 1:500), and anti‑phospho-Syk (pSyk) (Cell Signaling Technology; cat. no. 2710 1:1000), anti‑DDEF1 (Santa cruz; cat. sc-374410 1:1000), anti‑NF-κB (Santa cruz; cat. sc-8008 1:1000) and anti‑β‑actin (Millipore Sigma; cat. no. A5441 1:2000).. Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.Gel electrophoresis was performed using PowerPac™ HC High‑Current Power Supply (1645052; Bio‑Rad), with a 10% gel used for all antibodies.



    Similar Products

    anti psyk  (Cell Signaling Technology Inc)
    95
    Cell Signaling Technology Inc anti psyk
    Anti Psyk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+phospho+syk+psyk/Phospho-Syk+(Tyr525%2F526)+Rabbit+mAb/pmc12913675-361-15-16
    Average 95 stars, based on 1 article reviews
    anti psyk - by Bioz Stars, 2026-09
    95/100 stars
    		  Buy from Supplier
    anti psyk tyr525 526  (Cell Signaling Technology Inc)
    93
    Cell Signaling Technology Inc anti psyk tyr525 526
    SYK enhances DC functions. (a) Western blot analysis of WT and SYK S544Y BMDC SYK phosphorylation <t>(Tyr525/526),</t> total SYK expression, JNK1 (T183/Y185) phosphorylation, total JNK1 expression, ERK1 phosphorylation (T203/Y205) and total ERK1 expression treated or not treated with R406 SYK inhibitor. ( n = 3) (b) MTS test of WT and SYK S544Y BMDCs in 24 and 48 h ( n = 3). (c) Percentage of CD11C + cells in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (d) Percentage of CD40 + , CD86 + , MHCII + cells of total CD11C + in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (e-f) Percentage and representative plots of mature FITC + BMDCs incubated at 4℃ (f) and 37 ℃ (g) for 1 h. ( n = 5). (g-h) Percentage and representative plots of immature FITC + BMDCs incubated in 4℃ (f) and 37 ℃ (g) for 1 h ( n = 5). (i) Proliferation of OT-II CD8 T cells cocultured with WT or SYK S544Y BMDCs in various ratios ( n = 3). (j) Representative plots show the proportion of proliferative CD8 T cells cocultured with WT or SYK S544Y BMDCs in 20:1, 10:1, and 5:1 ratios. (k) p-SYK expression in WT and SYK S544Y BMDCs after DC-T cell interaction by FACS ( n = 5). In all panels, NS was not significant, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. A p-value less than 0.05 was considered significant
    Anti Psyk Tyr525 526, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+phospho+syk+psyk/Phospho-Syk+(Tyr525%2F526)+Rabbit+mAb/pmc12827454-47-3-5
    Average 93 stars, based on 1 article reviews
    anti psyk tyr525 526 - by Bioz Stars, 2026-09
    93/100 stars
    		  Buy from Supplier
    psyk  (Cell Signaling Technology Inc)
    95
    Cell Signaling Technology Inc psyk
    SYK enhances DC functions. (a) Western blot analysis of WT and SYK S544Y BMDC SYK phosphorylation <t>(Tyr525/526),</t> total SYK expression, JNK1 (T183/Y185) phosphorylation, total JNK1 expression, ERK1 phosphorylation (T203/Y205) and total ERK1 expression treated or not treated with R406 SYK inhibitor. ( n = 3) (b) MTS test of WT and SYK S544Y BMDCs in 24 and 48 h ( n = 3). (c) Percentage of CD11C + cells in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (d) Percentage of CD40 + , CD86 + , MHCII + cells of total CD11C + in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (e-f) Percentage and representative plots of mature FITC + BMDCs incubated at 4℃ (f) and 37 ℃ (g) for 1 h. ( n = 5). (g-h) Percentage and representative plots of immature FITC + BMDCs incubated in 4℃ (f) and 37 ℃ (g) for 1 h ( n = 5). (i) Proliferation of OT-II CD8 T cells cocultured with WT or SYK S544Y BMDCs in various ratios ( n = 3). (j) Representative plots show the proportion of proliferative CD8 T cells cocultured with WT or SYK S544Y BMDCs in 20:1, 10:1, and 5:1 ratios. (k) p-SYK expression in WT and SYK S544Y BMDCs after DC-T cell interaction by FACS ( n = 5). In all panels, NS was not significant, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. A p-value less than 0.05 was considered significant
    Psyk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+phospho+syk+psyk/Phospho-Zap-70+(Tyr319)%2FSyk+(Tyr352)+Rabbit+mAb/pmc12756361-132-29-32
    Average 95 stars, based on 1 article reviews
    psyk - by Bioz Stars, 2026-09
    95/100 stars
    		  Buy from Supplier
    rabbit anti psyk y352 65e4 id catalog  (Cell Signaling Technology Inc)
    95
    Cell Signaling Technology Inc rabbit anti psyk y352 65e4 id catalog
    (A-C) Representative confocal micrographs showing a single z slice (∼280 nm) of IFN-γ-primed Csk AS BMDMs stained for total Lyn (yellow) and pSyk <t>Y352</t> (cyan) after 3 min treatments with (A) medium only (untreated), (B) 3-IB-PP1, or (C) depleted zy-mosan (zym dep ). Fluorescence in the DAPI channel shows an internalized zym dep (magenta). Scale bars: 5 μm (D) Interaction area for pSyk Y352 clusters formed spontaneously during 3-IB-PP1 treatment and semi-contiguous interaction area per phagocytosed zym dep particle. Points are single-punctum values combined from 5 cells, shown with standard error of the mean (SEM). Significance (Sig.) in all panels assessed via nonparametric t test and Kolmogorov-Smirnov test: *** P = 0.0001. Inset: view of the lower range of 3-IB-PP1-induced pSyk clustering, showing the preponderance of nanoclusters within the distribution.
    Rabbit Anti Psyk Y352 65e4 Id Catalog, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+phospho+syk+psyk/Phospho-Zap-70+(Tyr319)%2FSyk+(Tyr352)+Rabbit+mAb/bio_rxiv__2025__04__16__648985-179-7-4
    Average 95 stars, based on 1 article reviews
    rabbit anti psyk y352 65e4 id catalog - by Bioz Stars, 2026-09
    95/100 stars
    		  Buy from Supplier
    anti psyk tyr525 526 rabbit monoclonal antibody  (Cell Signaling Technology Inc)
    94
    Cell Signaling Technology Inc anti psyk tyr525 526 rabbit monoclonal antibody
    (A-C) Representative confocal micrographs showing a single z slice (∼280 nm) of IFN-γ-primed Csk AS BMDMs stained for total Lyn (yellow) and pSyk <t>Y352</t> (cyan) after 3 min treatments with (A) medium only (untreated), (B) 3-IB-PP1, or (C) depleted zy-mosan (zym dep ). Fluorescence in the DAPI channel shows an internalized zym dep (magenta). Scale bars: 5 μm (D) Interaction area for pSyk Y352 clusters formed spontaneously during 3-IB-PP1 treatment and semi-contiguous interaction area per phagocytosed zym dep particle. Points are single-punctum values combined from 5 cells, shown with standard error of the mean (SEM). Significance (Sig.) in all panels assessed via nonparametric t test and Kolmogorov-Smirnov test: *** P = 0.0001. Inset: view of the lower range of 3-IB-PP1-induced pSyk clustering, showing the preponderance of nanoclusters within the distribution.
    Anti Psyk Tyr525 526 Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+phospho+syk+psyk/Phospho-Syk+(Tyr525%2F526)+Rabbit+mAb/pmc11997114-140-31-37
    Average 94 stars, based on 1 article reviews
    anti psyk tyr525 526 rabbit monoclonal antibody - by Bioz Stars, 2026-09
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    SYK enhances DC functions. (a) Western blot analysis of WT and SYK S544Y BMDC SYK phosphorylation (Tyr525/526), total SYK expression, JNK1 (T183/Y185) phosphorylation, total JNK1 expression, ERK1 phosphorylation (T203/Y205) and total ERK1 expression treated or not treated with R406 SYK inhibitor. ( n = 3) (b) MTS test of WT and SYK S544Y BMDCs in 24 and 48 h ( n = 3). (c) Percentage of CD11C + cells in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (d) Percentage of CD40 + , CD86 + , MHCII + cells of total CD11C + in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (e-f) Percentage and representative plots of mature FITC + BMDCs incubated at 4℃ (f) and 37 ℃ (g) for 1 h. ( n = 5). (g-h) Percentage and representative plots of immature FITC + BMDCs incubated in 4℃ (f) and 37 ℃ (g) for 1 h ( n = 5). (i) Proliferation of OT-II CD8 T cells cocultured with WT or SYK S544Y BMDCs in various ratios ( n = 3). (j) Representative plots show the proportion of proliferative CD8 T cells cocultured with WT or SYK S544Y BMDCs in 20:1, 10:1, and 5:1 ratios. (k) p-SYK expression in WT and SYK S544Y BMDCs after DC-T cell interaction by FACS ( n = 5). In all panels, NS was not significant, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. A p-value less than 0.05 was considered significant

    Journal: Inflammation

    Article Title: SYK Activation Enhances Dendritic Cell Functions in Spontaneous Rheumatoid Arthritis

    doi: 10.1007/s10753-025-02405-2

    Figure Lengend Snippet: SYK enhances DC functions. (a) Western blot analysis of WT and SYK S544Y BMDC SYK phosphorylation (Tyr525/526), total SYK expression, JNK1 (T183/Y185) phosphorylation, total JNK1 expression, ERK1 phosphorylation (T203/Y205) and total ERK1 expression treated or not treated with R406 SYK inhibitor. ( n = 3) (b) MTS test of WT and SYK S544Y BMDCs in 24 and 48 h ( n = 3). (c) Percentage of CD11C + cells in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (d) Percentage of CD40 + , CD86 + , MHCII + cells of total CD11C + in BMDC derived from WT and SYK S544Y mice ( n ≥ 5). (e-f) Percentage and representative plots of mature FITC + BMDCs incubated at 4℃ (f) and 37 ℃ (g) for 1 h. ( n = 5). (g-h) Percentage and representative plots of immature FITC + BMDCs incubated in 4℃ (f) and 37 ℃ (g) for 1 h ( n = 5). (i) Proliferation of OT-II CD8 T cells cocultured with WT or SYK S544Y BMDCs in various ratios ( n = 3). (j) Representative plots show the proportion of proliferative CD8 T cells cocultured with WT or SYK S544Y BMDCs in 20:1, 10:1, and 5:1 ratios. (k) p-SYK expression in WT and SYK S544Y BMDCs after DC-T cell interaction by FACS ( n = 5). In all panels, NS was not significant, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. A p-value less than 0.05 was considered significant

    Article Snippet: For intracellular staining, anti-pSYK (Tyr525/526) (Cell Signaling Technology, 4349 S), diluted in Perm/Wash Buffer (BD Biosciences), were applied to cells and allowed to incubate for 30 min. After the staining steps, cells were washed twice with Perm/Wash Buffer and once with FACS buffer.

    Techniques: Western Blot, Phospho-proteomics, Expressing, Derivative Assay, Incubation

    (A-C) Representative confocal micrographs showing a single z slice (∼280 nm) of IFN-γ-primed Csk AS BMDMs stained for total Lyn (yellow) and pSyk Y352 (cyan) after 3 min treatments with (A) medium only (untreated), (B) 3-IB-PP1, or (C) depleted zy-mosan (zym dep ). Fluorescence in the DAPI channel shows an internalized zym dep (magenta). Scale bars: 5 μm (D) Interaction area for pSyk Y352 clusters formed spontaneously during 3-IB-PP1 treatment and semi-contiguous interaction area per phagocytosed zym dep particle. Points are single-punctum values combined from 5 cells, shown with standard error of the mean (SEM). Significance (Sig.) in all panels assessed via nonparametric t test and Kolmogorov-Smirnov test: *** P = 0.0001. Inset: view of the lower range of 3-IB-PP1-induced pSyk clustering, showing the preponderance of nanoclusters within the distribution.

    Journal: bioRxiv

    Article Title: Steady-state phosphorylation of SHIP1 by Lyn restricts macrophage activation in the absence of a phagocytic synapse

    doi: 10.1101/2025.04.16.648985

    Figure Lengend Snippet: (A-C) Representative confocal micrographs showing a single z slice (∼280 nm) of IFN-γ-primed Csk AS BMDMs stained for total Lyn (yellow) and pSyk Y352 (cyan) after 3 min treatments with (A) medium only (untreated), (B) 3-IB-PP1, or (C) depleted zy-mosan (zym dep ). Fluorescence in the DAPI channel shows an internalized zym dep (magenta). Scale bars: 5 μm (D) Interaction area for pSyk Y352 clusters formed spontaneously during 3-IB-PP1 treatment and semi-contiguous interaction area per phagocytosed zym dep particle. Points are single-punctum values combined from 5 cells, shown with standard error of the mean (SEM). Significance (Sig.) in all panels assessed via nonparametric t test and Kolmogorov-Smirnov test: *** P = 0.0001. Inset: view of the lower range of 3-IB-PP1-induced pSyk clustering, showing the preponderance of nanoclusters within the distribution.

    Article Snippet: Antibodies were obtained from Cell Signaling Technologies (rabbit anti-pSyk Y352 65E4 ID/catalog # (#)2717, rabbit anti-pPLCγ Y1217 #3871, rabbit anti-pSHIP1 Y1020 #3941, rabbit anti-pSHP-1 Y564 D11G5 #8849, rabbit anti-pErk1/2 T202/Y204 D13.14.4e #4370, rabbit anti-pAkt S473 193H12 #4058, rabbit anti-pPI3K p85(Y458)/p55(Y199) #4228, mouse anti-β-Actin 8H10D10 #3700), Abcam (Cambridge, United Kingdom) (mouse anti-LynA+B Lyn01 ab1890), Santa Cruz (Dallas, TX, USA) (rabbit anti-LynA+B 44 #sc-15, goat anti-Hck M-28 #sc-1428), and LICOR (donkey anti-mouse IgG 800CW #925-32212, donkey anti-rabbit IgG 800CW #925-32213, donkey anti-mouse IgG 680LT #925-68022, donkey anti-goat IgG 680LT #926-32214 ) .

    Techniques: Staining, Fluorescence

    Densitometry quantification of protein species in immunoblotted lysates from IFN-γ-primed Csk AS BMDMs after 5 min treatment with 3-IB-PP1 or zym dep probed by immunoblot for (A) pSyk Y352 , (B) pPLCγ2 Y1217 , (C) pPI3K p85/p55 (D) immunoprecipitated Ras-GTP, (E) pErk1/2 Y202/T204 , and (F) pAkt S473 . Error: SEM; n=9-14 for 3-IB-PP1 and 11-15 for zym dep . Sig. assessed via non-parametric t test and Kolmogorov-Smirnov test: * P = 0.0216, *** P = 0.0007, ** P < 0.0001.

    Journal: bioRxiv

    Article Title: Steady-state phosphorylation of SHIP1 by Lyn restricts macrophage activation in the absence of a phagocytic synapse

    doi: 10.1101/2025.04.16.648985

    Figure Lengend Snippet: Densitometry quantification of protein species in immunoblotted lysates from IFN-γ-primed Csk AS BMDMs after 5 min treatment with 3-IB-PP1 or zym dep probed by immunoblot for (A) pSyk Y352 , (B) pPLCγ2 Y1217 , (C) pPI3K p85/p55 (D) immunoprecipitated Ras-GTP, (E) pErk1/2 Y202/T204 , and (F) pAkt S473 . Error: SEM; n=9-14 for 3-IB-PP1 and 11-15 for zym dep . Sig. assessed via non-parametric t test and Kolmogorov-Smirnov test: * P = 0.0216, *** P = 0.0007, ** P < 0.0001.

    Article Snippet: Antibodies were obtained from Cell Signaling Technologies (rabbit anti-pSyk Y352 65E4 ID/catalog # (#)2717, rabbit anti-pPLCγ Y1217 #3871, rabbit anti-pSHIP1 Y1020 #3941, rabbit anti-pSHP-1 Y564 D11G5 #8849, rabbit anti-pErk1/2 T202/Y204 D13.14.4e #4370, rabbit anti-pAkt S473 193H12 #4058, rabbit anti-pPI3K p85(Y458)/p55(Y199) #4228, mouse anti-β-Actin 8H10D10 #3700), Abcam (Cambridge, United Kingdom) (mouse anti-LynA+B Lyn01 ab1890), Santa Cruz (Dallas, TX, USA) (rabbit anti-LynA+B 44 #sc-15, goat anti-Hck M-28 #sc-1428), and LICOR (donkey anti-mouse IgG 800CW #925-32212, donkey anti-rabbit IgG 800CW #925-32213, donkey anti-mouse IgG 680LT #925-68022, donkey anti-goat IgG 680LT #926-32214 ) .

    Techniques: Western Blot, Immunoprecipitation

    (A-D) Immunoblots and quantifications of lysates from IFN-γ-primed Lyn +/+ and Lyn KO BMDMs probed for phosphorylated (A) Syk Y352 , (B) Erk1/2 T202/Y204 , (C) Akt S473 , and (D) SHIP1 Y1020 . β-Actin is a visual loading control, and quantifications were normalized to total protein content in the gel lane. Data are shown relative to the signal in Lyn +/+ . n=10-15 for Lyn +/+ and 6-7 for Lyn KO . Error bars: SEM. Sig. nonparametric, unpaired t with Kolmogorov-Smirnov test, *P=0.0152, **P≥0.007, ***P=0.0006 and ****P≤0.0001. (E) Representative images of PIP 3 (yellow) and nuclear staining (cyan) in IFN-γ -primed BMDMs. (F) Quantification of the geometric mean fluorescence intensity (gMFI) of PIP 3 staining in Lyn KO relative to Lyn +/+ BMDMs. Data points reflect the mean and SEM of 3-4 independent wells from 4 biological replicates. Sig. nonparametric, unpaired t test with Kolmogorov-Smirnov test, *** P<0.0006.

    Journal: bioRxiv

    Article Title: Steady-state phosphorylation of SHIP1 by Lyn restricts macrophage activation in the absence of a phagocytic synapse

    doi: 10.1101/2025.04.16.648985

    Figure Lengend Snippet: (A-D) Immunoblots and quantifications of lysates from IFN-γ-primed Lyn +/+ and Lyn KO BMDMs probed for phosphorylated (A) Syk Y352 , (B) Erk1/2 T202/Y204 , (C) Akt S473 , and (D) SHIP1 Y1020 . β-Actin is a visual loading control, and quantifications were normalized to total protein content in the gel lane. Data are shown relative to the signal in Lyn +/+ . n=10-15 for Lyn +/+ and 6-7 for Lyn KO . Error bars: SEM. Sig. nonparametric, unpaired t with Kolmogorov-Smirnov test, *P=0.0152, **P≥0.007, ***P=0.0006 and ****P≤0.0001. (E) Representative images of PIP 3 (yellow) and nuclear staining (cyan) in IFN-γ -primed BMDMs. (F) Quantification of the geometric mean fluorescence intensity (gMFI) of PIP 3 staining in Lyn KO relative to Lyn +/+ BMDMs. Data points reflect the mean and SEM of 3-4 independent wells from 4 biological replicates. Sig. nonparametric, unpaired t test with Kolmogorov-Smirnov test, *** P<0.0006.

    Article Snippet: Antibodies were obtained from Cell Signaling Technologies (rabbit anti-pSyk Y352 65E4 ID/catalog # (#)2717, rabbit anti-pPLCγ Y1217 #3871, rabbit anti-pSHIP1 Y1020 #3941, rabbit anti-pSHP-1 Y564 D11G5 #8849, rabbit anti-pErk1/2 T202/Y204 D13.14.4e #4370, rabbit anti-pAkt S473 193H12 #4058, rabbit anti-pPI3K p85(Y458)/p55(Y199) #4228, mouse anti-β-Actin 8H10D10 #3700), Abcam (Cambridge, United Kingdom) (mouse anti-LynA+B Lyn01 ab1890), Santa Cruz (Dallas, TX, USA) (rabbit anti-LynA+B 44 #sc-15, goat anti-Hck M-28 #sc-1428), and LICOR (donkey anti-mouse IgG 800CW #925-32212, donkey anti-rabbit IgG 800CW #925-32213, donkey anti-mouse IgG 680LT #925-68022, donkey anti-goat IgG 680LT #926-32214 ) .

    Techniques: Western Blot, Control, Staining, Fluorescence

    Immunoblots and quantifications of (A-B) pSyk Y352 , (C-D) pErk1/2 T202/Y204 , (E-F) pAkt S473 , and (G-H) pSHIP1 Y1020 in IFN-γ-primed Csk AS (Lyn +/+ ) or Csk AS Lyn KO (Lyn KO ) BMDMs treated with (A, C, E, G) 3-IB-PP1 or (B, D, F, H) zym dep , corrected for total protein in each gel lane and shown relative to t=0 of Lyn +/+ ; n=10-11 for Lyn +/+ , 4-7 for Lyn KO . Error bars: SEM. Sig. via two-way ANOVA with Sidak’s multiple comparisons test: * P = 0.0143, ** P = 0.0041, **** P<0.0001.

    Journal: bioRxiv

    Article Title: Steady-state phosphorylation of SHIP1 by Lyn restricts macrophage activation in the absence of a phagocytic synapse

    doi: 10.1101/2025.04.16.648985

    Figure Lengend Snippet: Immunoblots and quantifications of (A-B) pSyk Y352 , (C-D) pErk1/2 T202/Y204 , (E-F) pAkt S473 , and (G-H) pSHIP1 Y1020 in IFN-γ-primed Csk AS (Lyn +/+ ) or Csk AS Lyn KO (Lyn KO ) BMDMs treated with (A, C, E, G) 3-IB-PP1 or (B, D, F, H) zym dep , corrected for total protein in each gel lane and shown relative to t=0 of Lyn +/+ ; n=10-11 for Lyn +/+ , 4-7 for Lyn KO . Error bars: SEM. Sig. via two-way ANOVA with Sidak’s multiple comparisons test: * P = 0.0143, ** P = 0.0041, **** P<0.0001.

    Article Snippet: Antibodies were obtained from Cell Signaling Technologies (rabbit anti-pSyk Y352 65E4 ID/catalog # (#)2717, rabbit anti-pPLCγ Y1217 #3871, rabbit anti-pSHIP1 Y1020 #3941, rabbit anti-pSHP-1 Y564 D11G5 #8849, rabbit anti-pErk1/2 T202/Y204 D13.14.4e #4370, rabbit anti-pAkt S473 193H12 #4058, rabbit anti-pPI3K p85(Y458)/p55(Y199) #4228, mouse anti-β-Actin 8H10D10 #3700), Abcam (Cambridge, United Kingdom) (mouse anti-LynA+B Lyn01 ab1890), Santa Cruz (Dallas, TX, USA) (rabbit anti-LynA+B 44 #sc-15, goat anti-Hck M-28 #sc-1428), and LICOR (donkey anti-mouse IgG 800CW #925-32212, donkey anti-rabbit IgG 800CW #925-32213, donkey anti-mouse IgG 680LT #925-68022, donkey anti-goat IgG 680LT #926-32214 ) .

    Techniques: Western Blot

    Immunoblots and quantification of (A) pSHIP1 Y1020 , (B) pAkt S473 , (C) pSyk Y352 , and (D) pErk T202/Y204 in CskAS (Hck +/+ Fgr +/+ ) and Csk AS Hck KO Fgr KO (Hck KO Fgr KO ) BMDMs, corrected for the total protein in each lane and shown relative to t=0 of Hck +/+ Fgr +/+ . Error bars: SEM, n=10-11 for Hck +/+ Fgr +/+ and n=4 for Hck KO Fgr KO . Sig. 2-way ANOVA with Sidak’s multiple comparisons test: * P = 0.0499.

    Journal: bioRxiv

    Article Title: Steady-state phosphorylation of SHIP1 by Lyn restricts macrophage activation in the absence of a phagocytic synapse

    doi: 10.1101/2025.04.16.648985

    Figure Lengend Snippet: Immunoblots and quantification of (A) pSHIP1 Y1020 , (B) pAkt S473 , (C) pSyk Y352 , and (D) pErk T202/Y204 in CskAS (Hck +/+ Fgr +/+ ) and Csk AS Hck KO Fgr KO (Hck KO Fgr KO ) BMDMs, corrected for the total protein in each lane and shown relative to t=0 of Hck +/+ Fgr +/+ . Error bars: SEM, n=10-11 for Hck +/+ Fgr +/+ and n=4 for Hck KO Fgr KO . Sig. 2-way ANOVA with Sidak’s multiple comparisons test: * P = 0.0499.

    Article Snippet: Antibodies were obtained from Cell Signaling Technologies (rabbit anti-pSyk Y352 65E4 ID/catalog # (#)2717, rabbit anti-pPLCγ Y1217 #3871, rabbit anti-pSHIP1 Y1020 #3941, rabbit anti-pSHP-1 Y564 D11G5 #8849, rabbit anti-pErk1/2 T202/Y204 D13.14.4e #4370, rabbit anti-pAkt S473 193H12 #4058, rabbit anti-pPI3K p85(Y458)/p55(Y199) #4228, mouse anti-β-Actin 8H10D10 #3700), Abcam (Cambridge, United Kingdom) (mouse anti-LynA+B Lyn01 ab1890), Santa Cruz (Dallas, TX, USA) (rabbit anti-LynA+B 44 #sc-15, goat anti-Hck M-28 #sc-1428), and LICOR (donkey anti-mouse IgG 800CW #925-32212, donkey anti-rabbit IgG 800CW #925-32213, donkey anti-mouse IgG 680LT #925-68022, donkey anti-goat IgG 680LT #926-32214 ) .

    Techniques: Western Blot